TW201724969A - Method for manufacturing plural enzymes product with high activity - Google Patents

Abstract

The present invention discloses a method for manufacturing a highly active plural enzymes product, comprising the steps of: (a) providing plural fermentation bacteria and fruit(s) and/or vegetable(s) jam in a container to form a mixing fermentation liquid; (b) the plural fermentation bacteria is fermented as injecting carbon dioxide, oxygen and nitrogen into the container, in a first fermentation stage; (c) after the first fermentation stage, a second fermentation stage is conducted; and (d) a vacuum ultrasonic and homogenization treatment for the mixed fermentation liquid in the container is performed. During the fermentation, PH value, the proportion of carbon dioxide, oxygen, and nitrogen are adjusted as different stages.

Description

High-activity multi-enzyme product manufacturing method

The invention relates to an enzyme product, in particular to a method for producing a highly active multi-enzyme product.

Enzyme, also known as "enzyme", is essential for maintaining normal body functions, digesting food, and repairing tissues. Enzymes play a catalytic role and role in the human body. If the human body lacks enzymes, all activities in the body will stop. So, just have enough vitamins, minerals, water and protein, but without enzymes, you can't sustain life. The human body can synthesize enzymes from the pancreas and liver cells, but it can also be obtained from food. It is rich in enzymes in fresh fruits and vegetables and various vegetables.

Although the liver and pancreas of the human body can secrete enzymes by themselves, as the age increases, the ability to secrete enzymes will gradually decrease, and human enzymes will also be caused by diet, lifestyle, pollution, stress, smoking, drinking, etc. Loss. Therefore, it needs to be taken from daily food, but most of the enzymes will reduce the activity of the enzyme and lose its effect after cooking and heating. Therefore, it is necessary to supplement the enzyme with external food to keep enough enzymes in the body to ensure health and disease.

The important biochemical and metabolic reactions in the human body are very complex, and the reaction process can be carried out step by step, mainly relying on the role of dozens of enzymes. Enzymes are important components of the body's biochemical metabolism and maintenance of physiological functions. They are widely found in natural plants and living animals. The enzyme itself is a protein structure, which is a function of inhibiting bacteria, sterilizing, decomposing waste, removing phytotoxicity and pollution. Every day, the human body has weakened or damaged cells, so it strengthens the enzymes produced in the body or ingests fresh enzymes to provide the enzymes needed by the cells.

Enzyme is a catalyst for the human body. It is composed of a variety of amino acids and is a physiologically active protein. Enzymes belong to a protein and are therefore sensitive to heat. The activity of the enzyme increases with increasing temperature, but when the temperature reaches above 45 °C, its enzyme activity begins to decrease with increasing temperature. Enzymes are specific, and each enzyme can only catalyze a biochemical reaction.

The nutrients absorbed by the human body do not decompose, form cells and synthesize related nutrients. It is necessary to promote metabolism through enzymes. There are many natural enzymes in human cell tissues to complete the biochemical functions of tissues and organs. For the human body, in addition to the self-synthesizing enzyme, it is necessary to take the active enzyme in the natural food to maintain the body's metabolism, physiological and biochemical functions and promote health.

However, most of the foods ingested by the human body are processed products. The enzymes contained in the human body enter the stomach (digestive tract) of the human body. The protein is denatured due to the action of acid and alkali, and the structure changes, so that the enzyme activity is destroyed and lost. Original function.

Enzyme food is based on the addition of microbial strains and sugar to fruits and vegetables for fermentation. Generally, the types of fruits and vegetables, fermentation strains, and fermentation days used in the self-brewed enzyme solution are relatively small and short, and the temperature, pH, and fermentation changes of the whole fermentation process are difficult to control, so that the fermentation is carried out. The types and amounts of nutrients and enzymes are worse than the enzyme foods produced by professional biotechnology companies, and the quality is also unstable. Usually, high-quality enzyme foods need to be fermented for at least 540 days in order to ferment all kinds of beneficial substances that are most suitable for the human body. However, making enzyme foods for such a long time does not meet economic costs and benefits.

The method for preparing the high-activity multi-enzyme product proposed by the invention can balance the economic cost and the quality of the product.

In the present invention, a method for producing a high-activity multi-enzyme product comprises: providing a plurality of fermentation strains and pulp in a container to form a mixed fermentation liquid; and then containing a plurality of fermentation strains and fruits The container of the mixed fermentation broth is filled with carbon dioxide, oxygen and nitrogen, and is fermented in a first stage, wherein the pH is controlled between 5.0 and 6.5, the proportion of carbon dioxide is about 2 to 4%, and the proportion of oxygen is about 18~22%, the ratio of nitrogen is about 76~80%; after that, after the first stage, a second stage fermentation is carried out, wherein the pH is controlled between 3.0 and 4.5, and the proportion of carbon dioxide is about 2~5%. The ratio of oxygen is about 10 to 15%, and the ratio of nitrogen is about 80 to 88%. After the second stage, a mixed fermentation liquid in a vacuum ultrasonic homogenization treatment vessel is performed, wherein the pH is controlled between ~ .

In particular, the plurality of fermentation strains described above comprise a plurality of anaerobic strains, a plurality of aerobic strains, and a plurality of facultative strains. In addition, the plurality of fermentation strains include Streptococcus thermophilus, Lactobacillus paracasei, Lactobacillus bulgaricus, Bifidobacterium A, Bifidobacterium B, yeast, vinegar Acid bacteria, Kefir lactic acid bacteria, Rhizopus, R. rhamnosus, Lactobacillus fermentum, Lactobacillus helveticus, and Bacillus coagulans.

Furthermore, the above vacuum ultrasonic homogenization treatment tank mixes the fermentation broth, wherein the pH is controlled between 5.0 and 6.5. The vacuum ultrasonic homogenization process is a vacuum ultrasonic homogenization synchronous process.

These and other advantages are apparent from the following description of the preferred embodiments and claims.

The invention is described in detail herein with reference to the particular embodiments of the invention, and the description of the invention. Therefore, the present invention may be widely practiced in other different embodiments in addition to the specific embodiments and preferred embodiments of the specification.

An embodiment described in the specification refers to a particular feature, method or characteristic described in connection with this embodiment, which is included in at least some embodiments. Therefore, the implementation of the various embodiments or aspects of the various embodiments is not necessarily the same embodiment. Furthermore, the features, methods, or characteristics of the invention may be combined as appropriate in one or more embodiments.

It is worth noting that the fermentation strain provided by the present invention can be applied to any enzyme product. The enzyme product of the present invention is produced using 13 strains of fermented bacteria.

In one embodiment, the enzyme food of the present invention is a vegetable and fruit added to a microbial strain and sugar for fermentation. The type of fruits and vegetables used in the brewed enzyme solution, the type of fermentation strain, and the number of fermentation days are controlled, and the temperature, pH value, and proportion of ventilation gas in the whole fermentation process must also be controlled in order to control the fermentation change of the strain. Get the quality of the best enzyme products.

In general, the main gases that affect the growth and metabolism of fermenting species are oxygen and carbon dioxide. According to the different degrees of oxygen demand, the fermentation strains may include: 1. Those who need sufficient oxygen are aerobic bacteria, such as Pseudomonas, Artococcus, Acetococcus, etc.; Oxygen bacteria, such as Clostridium; Third, aerobic or non-oxygen can survive the facultative bacteria.

The cells of the aerobic bacteria are subjected to respiratory metabolism, the inner membrane of the cell contains an electronic transfer chain of integrity, and oxygen is the recipient of the final electron in the chain. General enzymes, such as glucose, undergo electron transfer After the metabolism of the chain, the final product is carbon dioxide and water. The cells of the anaerobic bacteria ferment the enzyme in an anaerobic state, and the final product is still an organic substance, which means that the enzyme is not completely utilized. The facultative bacteria are subjected to respiratory metabolism under aerobic conditions, and the fermentation reaction is carried out under anaerobic conditions. Under aerobic conditions, the facultative bacteria grow more and the growth rate is faster. Therefore, aerobic bacteria are more efficient at using enzymes than anaerobic bacteria. In the aerobic state, peroxide or hydrogen peroxide is often produced in the metabolism of cells. Aerobic bacteria, containing catalase or peroxide catalyzing enzymes, can decompose peroxides or hydrogen peroxide into non-toxic products. There is no such enzyme that decomposes peroxide in anaerobic bacteria. Therefore, in anaerobic bacteria, growth is inhibited or even killed.

In a container (equipment) containing an enzyme solution, the aerobic bacteria appear on the upper layer of the liquid interface due to the need for oxygen, the anaerobic bacteria appear at the bottom of the container without oxygen, and the anaerobic bacteria can be dispersed throughout the liquid interface. (Apparently between the upper and the bottom).

In the present invention, a method for producing a highly active multi-enzyme product includes the following several steps.

Step 1: providing a series of (plural) fermentation strains, fruits and vegetables in a container to form a fermentation liquid in which the fruits and vegetables can be pre-placed. The container is a fermenter.

In this step, the enzyme material is washed, broken, and beaten to form a pulp. The enzyme raw material can be vegetables or fruits. The main raw materials of the enzyme raw materials are, for example, pineapple, papaya, apple, grape, boluo, etc., and the auxiliary materials are, for example, yellow plum pulp, raisins, and the like. The type and weight ratio of the main raw materials and auxiliary raw materials can be selected and determined as needed. After the fruit pulp is formed, the bacteria are collected, and the pulp is mixed with the strain according to a certain ratio, and then placed in the fermentation tank to form a fermentation liquid.

Fermentation species include the following components: (a) an anaerobic strain; (b) an aerobic strain; and (c) a facultative aerobes. The facultative bacteria are microorganisms that can perform both aerobic respiration and anaerobic respiration or fermentation. When oxygen is sufficient, they produce ATP (adenosine triphosphate) by aerobic respiration, but when oxygen is deficient, their breathing patterns become anaerobic respiration. Aerobic bacteria (obligate aerobic microorganisms) cannot produce ATP in an anaerobic environment, while anaerobic bacteria (obligate anaerobic microorganisms) die in an aerobic environment because there is no substance such as catalase. For example, facultative bacteria include Staphylococcus, Streptococcus, Listeria, and Shewana Onida. Yeast is a typical facultative bacterium. Microorganisms such as facultative bacteria have a wide range of adaptations and can grow in aerobic or anaerobic environments. Generally, it is mainly aerobic growth, and it is dependent on respiratory productivity when aerobic; it has both anaerobic growth ability and fermentative or anaerobic respiration capacity when there is no oxygen. For example, Bacillus lichenifornus, Saccharomyces cerevisiae, and the like.

In one embodiment, in step 1, the fermenting species includes the following 13 strains: Streptococcus thermophilus, Lactobacillus paracasei, Lactobacillus bulgaricus, Bifi Lactobacillus acidophilus, Bifidobacterium, Yeast, Acetic acid bacillus, Kefir acidophilus, Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus fermentum, Lactobacillus helvetics, and Bacillus coagulans.

In the first step, the above 13 kinds of fermentation strains are mixed with the pulp to form a fermentation liquid, and three layers are automatically formed, wherein the aerobic bacteria appear in the upper layer of the fermentation liquid due to the need of oxygen, and the anaerobic bacteria do not need oxygen. At the bottom of the container, the facultative bacteria appear roughly between the upper layer and the bottom.

Step 2: Fermenting in a first stage by introducing carbon dioxide, oxygen and nitrogen into a container containing 13 kinds of fermentation broth and fermented fermentation broth.

In the first stage of fermentation (initial fermentation), the temperature is controlled at 20 to 30 ° C, and the pH is controlled between 5.0 and 6.5. Among the gases introduced, the ratio of carbon dioxide is about 2 to 4%, the ratio of oxygen is about 18 to 22%, and the ratio of nitrogen is about 76 to 80% to control the fermentation of the strain. In the first phase, the number of fermentation days was approximately controlled at 10 days.

Step 3: Fermenting in a second stage by introducing carbon dioxide, oxygen and nitrogen into a container containing 13 kinds of fermentation broth and fermented fermentation broth.

A step of adding sugar after the fermentation in the first stage and before the fermentation in the second stage, for example, adding an appropriate amount of sucrose, xylitol sugar, fructooligosaccharide and/or maltose, etc., is included.

In the second stage fermentation (post-fermentation), the temperature is controlled at 20 to 30 ° C, and the pH is controlled between 3.0 and 4.5. Among the gases introduced, the ratio of carbon dioxide is about 2 to 5%, the ratio of oxygen is about 10 to 15%, and the ratio of nitrogen is about 80 to 88% to control the fermentation of the strain. In the second stage, the number of fermentation days was approximately controlled at 20 days.

Step 4: Carbon dioxide, oxygen and nitrogen are introduced into the container containing the mixed fermentation broth of the fermenting bacteria and the pulp, and the mixed fermentation broth in the container is homogenized by a vacuum ultrasonic wave.

Vacuum ultrasonic homogenization includes ultrasonic and homogeneous synchronization, or ultrasonic and homogeneous non-synchronous processing. Ultrasonic and homogeneous synchronization processing means that ultrasonic processing and homogenization are the same. The time period is processed, and the ultrasonic and homogeneous asynchronous processing means that the ultrasonic processing and the homogenizing processing are processed at different time periods.

In the vacuum ultrasonic homogenization process, the temperature is controlled at 5~15 °C, and the PH value is controlled between 5.0~6.5. During this phase, the time for ultrasonic homogenization is approximately 20 to 30 minutes. The use of vacuum ultrasonic homogenization can quickly disintegrate bacterial cell bodies (cell wall rupture) and release enzymes.

The table below shows the growth of various microorganisms during the fermentation of fruits and vegetables. Taking four kinds of microorganisms as an example, they are synchronized with vacuum ultrasonic homogenization at the initial stage of fermentation (the first stage mentioned above) and the late stage of fermentation (the second stage mentioned above). Microbial growth data for treatment in CFU/ml. Colony-Forming Units (CFU) refer to the total number of bacterial communities per unit volume. The results show that aerated microbes still grow faster than non-ventilated microorganisms.

Microbial species A includes Streptococcus thermophilus, and microbial species B includes Lactobacillus paracasei, Lactobacillus bulgaricus, Bifidobacterium A, Bifidobacterium B, Kefir lactic acid bacteria, Rhizopus, R. rhamnosus, Lactobacillus fermentum, Lactobacillus helveticus, and Bacillus licheniformis, the microbial species C includes yeast, and the microbial species D includes acetic acid bacteria.

In general, digestive enzymes are generally divided into four types: 1. amylolytic enzymes, specifically decomposing starch; 2. protein decomposing enzymes, responsible for decomposing proteins; 3. lipolytic enzymes, responsible for breaking down fat; 4. fibrinolytic enzymes, responsible for Decompose the fiber. Immature papaya and pineapple are excellent sources of digestive enzymes (papaya enzyme, pineapple enzyme). Table 2 shows the activity of various enzymes during the fermentation of the fruit and vegetable enzymes. The five enzymes are exemplified by the enzymes in the early stage of fermentation (the first stage mentioned above) and the late stage of fermentation (the second stage mentioned above) and the simultaneous treatment with vacuum ultrasonic homogenization. Activity in U/g (mL) / min. The International Unit for Enzyme Activity (IU) is defined as the amount of enzyme required to catalyze the production of 1 μmol of product per minute.

Form 2

In the above embodiments, the enzyme prepared according to the method of the present invention is subjected to an enzyme activity change test in the initial stage of fermentation, the late fermentation stage and the ultrasonic treatment, and the test results are shown in Table II. . As can be seen from Table 2, (1) in the first stage of the initial fermentation, the enzyme activities in the five enzymes are: proteolytic enzyme, amylolytic enzyme, lipolytic enzyme, fibrinolytic enzyme, super oxidase (2) In the second stage of the late fermentation stage, there are two differences between aeration and non-ventilation. The activity of the aerated enzyme is about 0.5 to 1 times that of the unventilated enzyme. The enzyme activity of the five enzymes is higher than that of the early fermentation. The growth is in the order of (with ventilation): amylolytic enzymes, proteolytic enzymes, super oxidase, lipolytic enzymes, fibrinolytic enzymes, in which the amylolytic enzymes grow several times and grow the most; (3) After the simultaneous treatment of vacuum ultrasonic homogenization, the enzyme activities of the five decomposing enzymes have further growth compared with the late fermentation stage, and the size is (ventilation): amylolytic enzyme, proteolytic enzyme, super oxidase, lipid decomposition Enzymes, fibrinolytic enzymes, which still have the largest growth of amylolytic enzymes, about six times as much as the initial fermentation; (4) with gas (carbon dioxide) Oxygen and nitrogen) enzyme activity of about from about 1.5 to 2 times the enzyme activity does not pass gas (carbon dioxide, oxygen and nitrogen) of the. From this, it can be seen that the growth ratio of the enzyme activity is more pronounced after the gas is introduced and the vacuum ultrasonic homogenization is simultaneously performed.

As can be seen from the above, in the present invention, a mixed fermentation broth of a plurality of fermenting bacteria and fruit pulp can be appropriately added to the enzyme product, and after a certain number of control days, the temperature, the pH value, the introduction of a certain gas, and the like are controlled during the fermentation. The ratio of the gas and the simultaneous homogenization of the vacuum ultrasonic wave can control the fermentation changes of the strain to obtain the best quality of the enzyme product.

Advantages of the present invention include: (1) shortening of the production time of the enzyme product: preparation of the enzyme product can be completed in about 30 days; (2) the amount of the preservative can be reduced by the enzyme product of the present invention; (3) utilization of the present invention Fermented enzyme product produced by introducing gas into the fermentation tank The activity of the enzyme can be greatly improved; (4) the mixed fermentation broth of the present invention which utilizes vacuum ultrasonic homogenization and simultaneous treatment in the fermentation tank, and the enzyme activity of the produced enzyme product can be greatly improved; (5) The enzyme of the present invention The product can reduce the damage of the pH to the enzyme.

An embodiment is an implementation or example of the invention. An embodiment, an embodiment, some embodiments, or other embodiments described in the specification are specific to the features, structures, or characteristics described in connection with the embodiments, but not necessarily in some embodiments, but not necessarily In all embodiments. The embodiments of the various aspects are not necessarily the same embodiment. It should be understood that in the description of the embodiments of the present invention, the features are sometimes combined in the drawings and text descriptions of an embodiment, the purpose of which is to simplify the technical features of the present invention, and to facilitate understanding of the implementation of various aspects of the present invention. the way. In addition, the various modifications that can be made by the embodiments and the embodiments described in the present invention are intended to be included within the scope of the present invention. Therefore, the drawings and the examples are intended to be illustrative and not to limit the invention, and the scope of the invention is intended to be limited only by the scope of the claims.

Claims (10)

A method for preparing a high-activity multi-enzyme product comprises: providing a plurality of fermentation strains and pulp in a container to form a mixed fermentation liquid; and introducing carbon dioxide and oxygen into the container containing the mixed fermentation liquid Nitrogen, fermented in a first stage; after the first stage, carbon dioxide, oxygen and nitrogen are introduced into the vessel containing the mixed fermentation broth for a second stage fermentation; and after the second stage And performing a vacuum ultrasonic homogenization treatment of the mixed fermentation liquid in the container. The method for preparing a high-activity multi-enzyme product according to claim 1, wherein the fermentation is carried out in the first stage, wherein the pH is controlled between 5.0 and 6.5, and the ratio of the carbon dioxide is about 2 to 4%, and the ratio of the oxygen is It is about 18~22%, and the ratio of nitrogen is about 76~80%. The method for producing a high-activity multi-enzyme product according to claim 1 or 2, wherein the fermentation is carried out in the second stage, wherein the pH is controlled between 3.0 and 4.5, and the ratio of the carbon dioxide is about 2 to 5%, the oxygen. The ratio is about 10 to 15%, and the ratio of the nitrogen is about 80 to 88%. The method for producing a high activity multi-enzyme product according to claim 1, wherein the vacuum ultrasonic homogenization treatment is a vacuum ultrasonic homogenization synchronization treatment. The method for producing a high-activity multi-enzyme product according to claim 1, wherein the plurality of fermentation strains include Streptococcus thermophilus, Lactobacillus paracasei, Lactobacillus bulgaricus, Bifidobacterium A, Biefeld Bacteria B, yeast, acetic acid bacteria, Kefir lactic acid bacteria, Rhizopus, R. rhamnosus, Lactobacillus fermentum, Lactobacillus helveticus, and Bacillus coagulans. A method for preparing a high-activity multi-enzyme product comprises: providing a plurality of fermentation strains and pulp in a container to form a mixed fermentation liquid; and introducing carbon dioxide and oxygen into the container containing the mixed fermentation liquid Nitrogen, fermented in a first stage; After the first stage, carbon dioxide, oxygen and nitrogen are introduced into the vessel containing the mixed fermentation broth for a second stage fermentation. The method for preparing a high-activity multi-enzyme product according to claim 6, wherein the fermentation is carried out in the first stage, wherein the pH is controlled between 5.0 and 6.5, and the proportion of the carbon dioxide is about 2 to 4%, and the proportion of the oxygen is It is about 18~22%, and the ratio of nitrogen is about 76~80%. The method for producing a high-activity multi-enzyme product according to claim 6 or 7, wherein the fermentation is carried out in the second stage, wherein the pH is controlled between 3.0 and 4.5, and the ratio of the carbon dioxide is about 2 to 5%, the oxygen. The ratio is about 10 to 15%, and the ratio of the nitrogen is about 80 to 88%. A method for producing a high-activity multi-enzyme product, comprising: providing a plurality of fermentation strains and pulp in a container to form a mixed fermentation liquid; performing a first stage in the container containing the mixed fermentation liquid Fermentation, wherein the pH is controlled between 5.0 and 6.5; after the first stage, a second stage fermentation is carried out in the vessel containing the mixed fermentation broth, wherein the pH is controlled between 3.0 and 4.5; After the second stage, a vacuum ultrasonication is performed to homogenize the mixed fermentation broth in the vessel. The method for producing a high activity multi-enzyme product according to claim 9, wherein the vacuum ultrasonic homogenization treatment is a vacuum ultrasonic homogenization synchronization treatment.